Abstract
Many active compounds may be excluded from biological assays due to their low aqueous solubility. In this study, a simple method for the determination of the solubility of compounds containing aromatic rings is proposed. In addition to DMSO, five organic solvents for screening experiments of TNF-α inhibitors were explored. DMSO and PEG3350 were the most suitable for both protein stability and ligand-binding experiments. In addition, glycerol is a promising solvent for the screening of other compounds for which it might provide acceptable solubilization, due to its strong tendency to preserve the protein. Moreover, a fluorescence binding assay was developed using the TNF-α/SPD304 system, and a Kd of 5.36 ± 0.21 μM was determined. The results of this study could be used for the future screening of potential TNF-α inhibitors, while the protocols developed in this work could be applied to other proteins.
Original language | English |
---|---|
Pages (from-to) | 137-141 |
Number of pages | 5 |
Journal | ACS Medicinal Chemistry Letters |
Volume | 4 |
Issue number | 1 |
DOIs | |
Publication status | Published - 10 Jan 2013 |
Keywords
- aqueous solubility
- fluorescence binding assay
- insoluble ligands
- protein stability
- solvent selection